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[急性脊髓损伤大鼠血清与脊髓的代谢谱研究 ¹H 核磁共振波谱法]

[The metabolic profilings study of serum and spinal cord from acute spinal cord injury rats ¹H NMR spectroscopy].

作者信息

Hu Hua-Hui, Huang Xiao-Long, Quan Ren-Fu, Yang Zong-Bao, Xu Jing-Jing

机构信息

Xiaoshan Hospital of Traditional Chinese Medicine, Hangzhou 311201, Zhejiang, China;

出版信息

Zhongguo Gu Shang. 2017 Feb 25;30(2):152-158. doi: 10.3969/j.issn.1003-0034.2017.02.012.

Abstract

OBJECTIVE

To establish the rat model of acute spinal cord injury, followed by aprimary study on this model with ¹H NMR based on metabonomics and to explore the metabonomics and biomarkers of spinal cord injury rat.

METHODS

Twenty eight-week-old adult male SD rats of clean grade, with body weight of (200±10) g, were divided into sham operation group and model group in accordance with the law of random numbers, and every group had 10 rats. The rats of sham operation group were operated without damaging the spinal cord, and rats of model group were made an animal model of spinal cord incomplete injury according to the modified Allen's method. According to BBB score to observate the motor function of rats on the 1th, 5th, and 7th days after surgery. Postoperative spinal cord tissue was collected in order to pathologic observation at the 7th day, and the metabolic profilings of serum and spinal cord from spinal cord injury rats were studied by ¹H NMR spectroscopy.

RESULTS

The hindlimb motion of rats did not obviously change in sham operation group, there was no significant difference at each time point;and rats of model group occurred flaccid paralysis of both lower extremities, there was a significant difference at each time; there was significant differences between two groups at each time. Pathological results showed the spinal cord structure was normal with uniform innervation in shame group, while in model group, the spinal cord structure was mussy, and the neurons were decreased, with inflammatory cells and necrotic tissue. Analysis of metabonomics showed that concentration of very low density fat protein (VLDL), low density fat protein (LDL), glutamine, citric acid, dimethylglycine (DMG) in the serum and glutathione, 3-OH-butyrate, N-Acetyl-L-aspartic acid (NAA), glycerophosphocholine (GPC), glutamic acid, and ascorbate in spinal cord had significant changes(<0.05).

CONCLUSIONS

There are significant differences in metabolic profile from serum and spinal cord sample between model group and sham operation group, it conduces to explain the changes of small molecular substances in serum and spinal cord tissue after spinal cord injury, this provides the research basis for targeted research on the role of metabolic markers in patients with acute spinal cord injury.

摘要

目的

建立大鼠急性脊髓损伤模型,并基于代谢组学对该模型进行¹H NMR初步研究,探索脊髓损伤大鼠的代谢组学特征及生物标志物。

方法

选取28只清洁级8周龄成年雄性SD大鼠,体重(200±10)g,按随机数字法分为假手术组和模型组,每组10只。假手术组大鼠手术时不损伤脊髓,模型组大鼠按改良Allen法制作脊髓不完全损伤动物模型。术后第1、5、7天根据BBB评分观察大鼠运动功能。术后第7天取脊髓组织进行病理观察,采用¹H NMR光谱法研究脊髓损伤大鼠血清和脊髓的代谢轮廓。

结果

假手术组大鼠后肢运动无明显变化,各时间点差异无统计学意义;模型组大鼠双下肢弛缓性瘫痪,各时间点差异有统计学意义;两组各时间点比较差异有统计学意义。病理结果显示,假手术组脊髓结构正常,神经支配均匀,而模型组脊髓结构紊乱,神经元减少,伴有炎性细胞和坏死组织。代谢组学分析显示,血清中极低密度脂蛋白(VLDL)、低密度脂蛋白(LDL)、谷氨酰胺、柠檬酸、二甲基甘氨酸(DMG)以及脊髓中谷胱甘肽、3-羟基丁酸、N-乙酰-L-天冬氨酸(NAA)、甘油磷酸胆碱(GPC)、谷氨酸和抗坏血酸的浓度有显著变化(<0.05)。

结论

模型组与假手术组血清和脊髓样本的代谢轮廓存在显著差异,有助于解释脊髓损伤后血清和脊髓组织中小分子物质的变化,为针对性研究代谢标志物在急性脊髓损伤患者中的作用提供了研究依据。

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